MLV-10A1 retrovirus pseudotype efficiently transduces primary human CD4(+)T lymphocytes

Background Previously, we showed that retroviral vectors pseudotyped with t he envelope of the amphotropic murine leukemia virus 10A1 (MLV10A1) more ef ficiently transduce primary human CD8(+) T lymphocytes when compared with o ther A-MLV, gibbon ape leukemia virus (GaLV) and feline endogenous retrovir us (RD114) vector pseudotypes. For the success of several gene therapeutic approaches (ADA, HIV) it is important to effectively transduce primary huma n CD4(+) T lymphocytes. Methods We have used retroviral vectors encoding the enhanced green fluores cent protein (EGFP) as a marker gene and carrying envelopes of MLV10A1, A-M LV and GaLV and have analyzed the transduction efficiency of both human CD4 (+) T cell lines (CEM, H9, HUT78, J16) and primary human CD4(+) T lymphocyt es using a RetroNectin-assisted transduction protocol and virus-containing supernatant Results In CD4(+) T cell lines the MLV-10A1 vector pseudotype was most effe ctive and infected up to 85% of cells which then stably expressed GFP over time. MLV-10A1 was also superior and infected approximately 32% of primary human CD4+ T lymphocytes in comparison to GaLV (18%) and A-MLV (12%). The s uperior efficiency of MLV-10A1 for the transduction of CD4(+) T cells corre lates with the longer half-life of this pseudotype in comparison to A-MLV a nd, as previously shown by interference analysis, with the usage of both th e A-MLV (Pit2) and the GaLV receptor (Pit1) for cell entry. Conclusions MLV-10A1 is a suitable vector for transferring genes with high efficacy into primary human CD4(+) T lymphocytes. The use of MLV-10A1 pseud otyped vectors should make it easier to obtain a sufficient number of gene- modified T lymphocytes for an adoptive transfer. Copyright (C) 2000 John Wi ley & Sons, Ltd.